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In vitro activity of minocycline against respiratory pathogens from patients with cystic fibrosis.

Our objective was to determine the in vitro activity of minocycline against isolates of Burkholderia cepacia (BC), Stenotrophomonas maltophilia (SM), and Pseudomonas aeruginosa (PA) cultured from the respiratory tract of patients with cystic fibrosis (CF). Cultures of BC, SM, and PA were isolated in a hospital bacteriology laboratory from the sputum or oropharyngeal cultures obtained from patients attending a Cystic Fibrosis Center, and were prospectively tested for in vitro sensitivity to minocycline by Kirby-Bauer disk diffusion. From January 1994 to July 1995, 116 cultures from 61 patients had at least one of the three pathogens; 9/61 (15%) patients had an isolate of BC, and 7/9 (78%) had an initial isolate sensitive to minocycline, of which 3 were sensitive only to minocycline; 2 cultures were resistant to all antibiotics. Four of 7 patients with BC were treated with minocycline; 3 patients developed resistant isolates 3-13 months after therapy. Five of 61 patients (8%) had an isolate of SM: 4/5 (80%) of these isolates were sensitive to minocycline, of which 1 was sensitive only to minocycline. Fifty-five of 61 patients (90%) had at least one PA isolate, with 112 morphotypes recovered from 90 cultures: 40/112 morphotypes (36%) were sensitive to minocycline, 65 (58%) were resistant, and 7 (6%) were intermediate in sensitivity. We conclude that the marked in vitro activity of minocycline against BC and SM isolated from patients with CF suggests that minocycline may have an adjunct role in the antimicrobial therapy of multidrug resistant, respiratory pathogens in CF.

Adolescent↗

Synthesis and antimicrobial activity of some 1,2,4-triazole-3-mercaptoacetic acid derivatives.

Ethyl 5-(2-furyl)-4-ethyl-1,2,4-triazole-3-mercaptoacetate (2), 5-(2-furyl)-4-ethyl-1,2,4-triazole-3-mercaptoacetic acid hydrazide (3) and a series of new N-alkylidene/arylidene-5-(2-furyl)-4-ethyl-1,2,4-triazole-3-mercaptoacetic acid hydrazides (4a-f) were synthesized and evaluated for in vitro antibacterial activity against Staphylococcus aureus ATCC 6538. Staphylococcus epidermidis ATCC 12228, Klebsiella pneumoniae ATCC 4352, Pseudomonas aeruginosa ATCC 1539, Escherichia coli ATCC 8739, Shigella flexneri, Salmonella typhi, Proteus mirabilis and antifungal activity against Candida albicans ATCC 10231 using the disk diffusion and microdilution methods. Compound 4f showed antibacterial activity against some bacteria. The in vitro antimycobacterial activity of the new compounds against Mycobacterium tuberculosis H37Rv was evaluated employing the BACTEC 460 radiometric system. The highest inhibition observed was 61% at > 6.25 microg/ml.

Anti-Bacterial Agents↗

Synthesis, degradation kinetics and in vitro antimicrobial activity of tetrahydro-2H-1,3,5-thiadiazine-2-thione derivatives of some beta-amino acids.

Two series of tetrahydro-2H-1,3,5-thiadiazine-2-thione (THTT) derivatives were synthesized, 2a-f and 3a-f, by incorporation of beta-alanine and beta-phenylalanine, respectively, at the 5th position of the THTT moiety. Structures of these derivatives were verified by spectral and elemental methods of analyses. The lipophilic properties of the synthesized derivatives, expressed as calculated log P (Clog P), revealed that the beta-phenylalanine derivatives, 3a-f, have enhanced lipophilic characters compared to the corresponding beta-alanine analogues, 2a-f. The stability of the synthesized derivatives in aqueous buffer solution of pH 7.4, and in 80% human plasma was studied at 37 degrees C using high pressure liquid chromatography. As a general pattern, under the investigation conditions beta-phenylalanine derivatives were more labile and susceptible for chemical and enzymatic degradation than the corresponding beta-alanine analogues. Furthermore, the degradation rates of the synthesized derivatives affected by the variation of substituents on N-3 of the THTT moiety. The anti-fungal activity of the synthesized compounds was tested in vitro against different strains of fungi using the standard agar disk diffusion method. beta-Alanine derivatives, 2e and 2f, bearing an aralkyl group on the 3ed position of the THTT moiety exhibited antifungal activity against C. albicans and F. oxysporum. Furthermore, 2a, which is also a beta-alanine derivative bearing an ethyi group on the 3ed position of the THTT moiety solely showed a significant in vitro anti-bacterial activity against Bacillus serreus (Gram positive) and Serratia rhodnii (Gram negative).

Alanine↗

Antimicrobial susceptibility pattern of Klebsiella species from Ebonyi State University Teaching Hospital Abakaliki, Nigeria.

Klesiella specie isolated from clinical specimens from Ebonyi State University Teaching Hospital (EBSUTH). Abakakliki were studied to determine the antimicrobial susceptibility pattern. Between January, 2003 and September 2004 a total of 3.600 specimens processed in the routine Medical Microbiology laboratory of EBSUTH, of which 245(6.8%) yielded Klebsiella species, with 84 from out - patients and 161 from in - patients. The number of isolates from various samples were: Urine 126, Sputum 37 Endocervical swab 13, Aspirates 8, High Vaginal Swab 7, Blood 3, Eye Swab, Ear Swab and Cerebrospinal fluid were 2 samples each. Organisms were identified by conventional methods. Antimicrobial susceptibility was done by the disk diffusion methods. The antimicrobial disk used include: Ceftazidime, Cefuroxime, Cefotaxine, Augmentin, Pefloxacin (30ug), Doxycyline (25ug) Genticin (10 ug) Ciprofloacin and Ofloxacin (5ug) each and Erythromycin (15ug). All were Oxoid products. Results were interpreted according to NCCLS criteria. Klebsilla species were isolated mostly from urine specimens (51.4%) followed by wound swabs (18.4%). Antimicrobial susceptibility to various groups drugs used was generally poor. The most sensitive antimicrobial was Ciprofloxacin with 121(49.4%) isolates susceptible to it, followed by Gentamicin with 95 (38.8%) and Ceftazidime with 90(36.7%). Seventeen isolates were multiresistant to all the antimicrobial agents used. The result of this study will help in the empiric therapy of infection caused by Klebsiella species in Ebonyi State University Teaching Hospital, Abakaliki, Nigeria but continuous surverillance of antimicrobial resistance of the organnisn is very necessary in the formulation of a sound antibiotic policy in the hospital.

Anti-Bacterial Agents↗

Antibiogram, biotyping and plasmid profile analysis of Acinetobacter calcoaceticus var. anitratus.

Antibiogram, biotyping, and plasmid profile analysis were studied on 115 isolates (40 nosocomial and 75 community strains) of Acinetobacter calcoaceticus var. anitratus isolated from clinical specimens. By disk diffusion method, we found 14 antimicrobial susceptibility patterns. Among them, the gentamicin-resistant patterns were recovered more frequently from nosocomial origin than from community-acquired origin. Fifteen patterns of biocodes were found by using GNI card of AutoMicrobic System (Vitek System). By using rapid alkaline extraction method and agarose gel electrophoresis, 102 isolates (88.7%) contained plasmids, and 72 distinct profiles were found. No particular plasmid profile correlates with specific antibiogram or biotype. However, multiple isolates with identical antibiogram and biotype recovered from the same patient were usually of identical plasmid profile. In view of the heterogeneity of the plasmid patterns that were found, the combined use of antibiogram, biotyping, and plasmid profile analysis could be satisfactory for epidemiological investigation.

Acinetobacter↗

Evaluation of subtherapeutic use of the antibiotics apramycin and carbadox on the prevalence of antimicrobial-resistant Salmonella infection in swine.

The antibiotics apramycin and carbadox were fed to growing swine, and the prevalence of Salmonella isolates that are resistant to apramycin and related aminoglycoside antibiotics was examined. Three hundred twelve Salmonella-positive pigs raised on one of five farms in an integrated swine operation and slaughtered at a central plant were used. All farms fed carbadox during the grower phase, and two farms administered apramycin during the first 21 days of age. Ileocolic lymph nodes and cecal contents were sampled at slaughter. One hundred of the 312 pigs were randomly selected to examine apramycin- and carbadox-resistant Salmonella infection, while all 312 pigs were used to evaluate the association between apramycin exposure and infection with Salmonella organisms resistant to amikacin, gentamicin, kanamycin, and streptomycin. Antimicrobial resistance was determined using disk diffusion and breakpoint concentrations. Apramycin treatment appeared to have little effect on apramycin- (12.5 versus 20.9%) or streptomycin- (76.4 versus 73.5%) resistant Salmonella isolates when averaged across farms and compared to control animals. Feeding carbadox resulted in carbadox-resistant Salmonella infection in only 5.3% of the isolates on one farm. The prevalence of amikacin-, gentamicin-, and kanamycin-resistant Salmonella isolates on farms feeding apramycin and carbadox were 0, 0, and 1.8%, respectively. Serogroup B was the most prevalent serogroup isolated, followed by C1 and E1. Apramycin and carbadox treatment did not appear to have any effect on the serogroup isolated. Subtherapeutic use of carbadox and apramycin did not appear to increase the prevalence of antimicrobial-resistant Salmonella in market-age swine.

Animals↗

RO 23-6240 (AM-833), a new fluoroquinolone: in vitro antimicrobial activity and tentative disk diffusion interpretive criteria.

The susceptibility of over 7000 recent clinical bacterial isolates to RO 23-6240, a new trifluorinated quinolone, was determined at four medical centers. Over 99% of Enterobacteriaceae and 97% of staphylococci were inhibited by less than or equal to 2.0 micrograms/ml of RO 23-6240. Only 71% of Pseudomonas spp. were inhibited by this concentration. Streptococci and enterococci were resistant to RO 23-6240. Clinical isolates of Haemophilus spp., pathogenic Neisseria spp., and Branhamella catarrhalis were inhibited by less than or equal to 0.25 micrograms/ml of RO 23-6240. This drug's antibacterial activity was comparable with that of enoxacin and norfloxacin, but was less than that of ciprofloxacin against most species. Using less than or equal to 2.0 micrograms/ml and greater than or equal to 8.0 micrograms/ml as the susceptible and resistant MIC breakpoints for RO 23-6240, the regression analysis-derived disk diffusion zone diameter breakpoints for the 5 micrograms disk are: Susceptible greater than or equal to 19 mm intermediate 16-18 mm, and resistant less than or equal to 15 mm.

Anti-Bacterial Agents↗

In vitro evaluation of cefixime (FK027, FR17027, CL284635): spectrum against recent clinical isolates, comparative antimicrobial activity, beta-lactamase stability, and preliminary susceptibility testing criteria.

Cefixime, a new orally absorbed cephalosporin, was compared by in vitro testing with other oral beta-lactams, including cephalexin, cefaclor, cefuroxime, amoxicillin, and amoxicillin + clavulanate. Enterobacteriaceae were inhibited by lower concentrations of cefixime than any of the reference drugs; 90% and 95% were inhibited by less than or equal to 1.0 and less than or equal to 8.0 micrograms/ml, respectively. Cefixime was the least active among these drugs against staphylococci, with only 31% of 1106 strains inhibited by less than or equal to 8.0 micrograms/ml and less than 1% by less than or equal to 1.0 microgram/ml. Enterococci and pseudomonads were not susceptible to any of the drugs tested. Penicillin-resistant pneumococci were relatively resistant to cefixime, but penicillin-susceptible pneumococci were very susceptible to cefixime. Other streptococci were generally susceptible to all compounds tested, with relative activities of amoxicillin greater than cefaclor and cefuroxime greater than cefixime greater than cephalexin. Cefixime was inactive against Bacteroides species. A slight inoculum effect occurred with cefixime with inocolum concentrations varying from 10(5) to 10(6) colony forming units per milliliter, but this was more marked at 10(7) colony forming units per milliliter. Cefixime was resistant to hydrolysis by seven common beta-lactamases. It inhibited the hydrolysis of nitrocefin only by type 1 cephalosporinases. The disk diffusion zone diameter breakpoints for the 30-micrograms cefixime disk were determined by regression analysis to be greater than or equal to 27 mm (susceptible) and less than or equal to 23 mm (resistant), respectively corresponding to minimal inhibitory concentration breakpoints of less than or equal to 1.0 and greater than or equal to 4.0 micrograms/ml. Because of the high interpretive error rate (13.8%) and the occurrence of these breakpoints on the parabolic portion of the regression curve, we recommend further evaluation of cefixime disks with lower potencies.

Amoxicillin↗

Susceptibility of Staphylococcus species and subspecies to teicoplanin.

Twenty-four Staphylococcus species and their subspecies were examined for their susceptibilities to teicoplanin by disk diffusion (30-micrograms disk) and agar dilution for the determination of MICs. Moderately susceptible and resistant clinical strains were further tested for their susceptibilities to oxacillin and vancomycin. Teicoplanin resistance was not observed in the reference strains of the various Staphylococcus species isolated from healthy volunteers or animals. However, the novobiocin-resistant species Staphylococcus saprophyticus, Staphylococcus cohnii, Staphylococcus xylosus, Staphylococcus arlettae, Staphylococcus kloosii, and Staphylococcus gallinarum were less susceptible to teicoplanin (MIC, 2 to 8 micrograms/ml) than most of the novobiocin-susceptible species were (MIC, 0.5 to 4 micrograms/ml). Clinical isolates of coagulase-negative species were generally less susceptible to teicoplanin than were reference strains. Seven percent of the Staphylococcus epidermidis clinical strains were moderately susceptible (MIC, 16 micrograms/ml) to teicoplanin. Of these strains, 70% were oxacillin resistant. For Staphylococcus haemolyticus strains, 11% were resistant (MIC, greater than 16 micrograms/ml) and 21% were moderately susceptible to teicoplanin. Of these strains, 95% were oxacillin resistant, No strains of S. epidermidis or S. haemolyticus were intermediate or resistant to vancomycin. Teicoplanin appears to be less active in vitro against oxacillin-resistant S. haemolyticus. However, teicoplanin is an effective antimicrobial agent against many Staphylococcus species.

Animals↗

Pharmacological properties of Moringa oleifera. 1: Preliminary screening for antimicrobial activity.

The antimicrobial activities of Moringa oleifera leaves, roots, bark and seeds were investigated in vitro against bacteria, yeast, dermatophytes and helminths pathogenic to man. By a disk-diffusion method, it was demonstrated that the fresh leaf juice and aqueous extracts from the seeds inhibit the growth of Pseudomonas aeruginosa and Staphylococcus aureus and that extraction temperatures above 56 degrees C inhibited this activity. No activity was demonstrated against four other pathogenic Gram-positive and Gram-negative bacteria and Candida albicans. By a dilution method, no activity was demonstrated against six pathogenic dermatophytes. A method was standardized for studying the effect of aqueous extracts on Ascaris lumbricoides eggs, but no activity was exhibited by any part of the tree in contrast to Chenopodium ambrosioides leaf extracts.

Animals↗

Lincomycin resistance in methicillin-resistant Staphylococcus aureus strains of hospital origin.

A total of 170 Staphylococcus aureus strains isolated during a one-year period at the University Hospital of Patras Medical School were examined for resistance to a battery of antimicrobial agents by disk diffusion and minimum inhibitory concentration (MIC) determination. Fifty-five isolates were lincomycin- and methicillin-resistant (LMRSA). In the group of 55 LMRSA isolates 13 were also resistant to vancomycin. All the LMRSA isolates were not typed by the international set and the experimental phages 88A and 25 at routine typing dilution (RTD), while 18 isolates were lysed by phages at 100XRTD and 1000XRTD. Reverse phage-typing and heat shock treatment of the LMRSA isolates had no effect on their typability. Plasmid profiles coupled with restriction endonuclease analysis of plasmid DNA established that the LMRSA isolates represent different strains. Membrane-protein profiles by polyacrylamide gel electrophoresis (PAGE) showed that LMRSA strains could belong to one group. This method proved useful and sensitive for characterization of LMRSA.

Bacterial Proteins↗

Meropenem susceptibility breakpoint for Pseudomonas aeruginosa strains hyperproducing mexB mRNA.

Twenty-five isolates of Pseudomonas aeruginosa with different meropenem susceptibilities were subjected to quantitative RT-PCR for analysis of transcription levels of oprD, mexB and mexD, and, in selected isolates, PA3720, which is hyper-expressed in nalC efflux mutants. Regulator genes of efflux pump MexAB-OprM, mexR and PA3721 (putative) were sequenced in selected isolates. The potential for mathematical reconstruction of the ideal susceptible population using normalised resistance interpretation (NRI) was also studied. In three isolates with intermediate susceptibility to meropenem (according to Swedish breakpoints), a reduction in MIC from 4 to 2 mg/L was observed with efflux inhibitor MC-207,110. These isolates would be considered susceptible according to British Society for Antimicrobial Chemotherapy and NCCLS breakpoints. These three isolates had between 4.6- and 5.0-fold increases in mexB transcription. None of these isolates had significant nalB mutations, but an Ala145-->Val mutation was observed in PA3721 in two of the isolates. However, these isolates had moderately increased production of PA3720 only. Single-strain regression analysis did not detect any major biological differences between the different groups. Using NRI, a disk-diffusion susceptibility breakpoint of >/= 28 mm was generated. Isolates with intermediate susceptibility to meropenem, which are considered fully susceptible in many countries, displayed possible low-grade meropenem resistance mechanisms, implying that the susceptibility breakpoint should be reconsidered. The increased transcription of mexB mRNA in such isolates seems unrelated to nalB or nalC mutations.

Anti-Bacterial Agents↗

The antimicrobial activities of Psidium guajava and Juglans regia leaf extracts to acne-developing organisms.

This study aims to present the in vitro inhibitory effect of Psidium guajava and Juglans regia leaf extracts on the main developer of acne lesions, Propionibacterium acnes (P. acnes), and other organisms that are isolated from acne lesions. Thirty-eight subjects (males and females) who had various types of acne were enrolled in the study. The contents of the acne lesions were cultured and the frequency of P. acnes (alone and with Staphylococci spp.) was 47%, whereas the frequencies for Staphylococcus aureus and Staphylococcus epidermidis were 13% and 24%, respectively. The antimicrobial activities of Psidium guajava and Juglans regia leaf extracts, determined by disk diffusion method (zone of inhibition), were compared to tea tree oil (TTO), doxycycline and clindamycin antibiotics. The zones of inhibition due to the Psidium guajava and Juglans regia leaf extracts ranged from 15.8-17.6 mm against P. acnes, 11.3-15.7 mm against S. aureus and 12.9-15.5 mm against S. epidermidis, respectively. These zones of inhibition were significantly higher than those of TTO and equivalent in case of Staphylococci spp., but less in case of P. acnes, to those obtained from doxycycline or clindamycin. It can be concluded that Psidium guajava and Juglans regia leaf extracts may be beneficial in treating acne especially when they are known to have anti-inflammatory activities.

Acne Vulgaris↗

Studies on the susceptibility of 150 consecutive clinical isolates of Pseudomonas aeruginosa to tobramycin, gentamicin, colistin, carbenicillin, and five other antimicrobials.

The in vitro susceptibilities of 150 clinical isolates of Pseudomonas aeruginosa to the aminoglycoside tobramycin and eight other antimicrobials were evaluated. The Food and Drug Administration standardized disk diffusion method showed that tobramycin inhibited 100% of the 150 isolates with gentamicin inhibiting only 90.7%. The difference between colistin (97.3%) and tobramycin was less marked. Carbenicillin (87.3%) was found to be slightly less active than gentamicin. Only a small percentage of the isolates were inhibited by the other drugs used. By using a minimal inhibitory concentration of 4 mug/ml in brain heart infusion broth as the level of susceptibility, it was found that only three of the isolates showed resistance to tobramycin, whereas 20 of the isolates were resistant to gentamicin. Approximately half of the isolates were inhibited by 0.5 mug/ml or less of tobramycin, whereas 1.5 mug of gentamicin per ml was required to inhibit 50% of susceptible strains.

Aminoglycosides↗

beta-Lactamases responsible for resistance to expanded-spectrum cephalosporins in Klebsiella pneumoniae, Escherichia coli, and Proteus mirabilis isolates recovered in South Africa.

Although resistance to the expanded-spectrum cephalosporins among members of the family Enterobacteriaceae lacking inducible beta-lactamases occurs virtually worldwide, little is known about this problem among isolates recovered in South Africa. Isolates of Klebsiella pneumoniae, Escherichia coli, and Proteus mirabilis resistant to expanded-spectrum cephalosporins recovered from patients in various parts of South Africa over a 3-month period were investigated for extended-spectrum beta-lactamase production. Antibiotic susceptibility was determined by standard disk diffusion and agar dilution procedures. Production of extended-spectrum beta-lactamases was evaluated by using the double-disk test, and the beta-lactamases were characterized by spectrophotometric hydrolysis assays and an isoelectric focusing overlay technique which simultaneously determined isoelectric points and general substrate or inhibitor characteristics. DNA amplification and sequencing were performed to confirm the identities of these enzymes. The P. mirabilis and E. coli isolates were found to produce TEM-26-type, SHV-2, and SHV-5 extended-spectrum beta-lactamases. An AmpC-related enzyme which had a pI of 8.0 and which conferred resistance to cefoxitin as well as the expanded-spectrum cephalosporins was found in a strain of K. pneumoniae. This is the first study which has identified organisms producing different extended-spectrum beta-lactamases from South Africa and the first report describing strains of P. mirabilis producing a TEM-26-type enzyme. The variety of extended-spectrum beta-lactamases found among members of the family Enterobacteriaceae isolated from major medical centers in South Africa is troubling and adds to the growing list of countries where these enzymes pose a serious problem for antimicrobial therapy.

Cephalosporin Resistance↗

Comparison of methods for the determination of antimicrobial resistance in Staphylococcus aureus from bovine mastitis.

The results of three standard methods (broth dilution, agar dilution, disk diffusion) and an experimental modification of the microdilution method for determination of resistance to ampicillin, cephalotin, cloxacillin, neomycin, novobiocin, penicillin and streptomycin were compared using 151 Staphylococcus aureus isolates obtained from cases of mastitis. The accuracy of the dilution methods was compared by determination of minimum inhibition concentrations (MIC, MIC50, MIC90 and modal MIC) and by assessment of the agreement within the tolerance of +/-1 dilution step in 2-fold dilution series. The results of the dilution methods were further compared with those of the reference disk diffusion method and the strains were classified as sensitive or resistant using the interpretation criteria for human strains. The comparisons indicated that MIC characteristics and the final classification as sensitive or resistant were method-dependent. Resistance to aminoglycoside antibiotics was observed more often when using broth dilution methods, especially when the broth was supplemented with lactose.

Ampicillin↗

Antimicrobial resistance phenotypes of ETEC isolates from piglets with diarrhea in North Vietnam.

Both disk diffusion and broth micro-dilution assays were employed to determine the level of resistance in Enterotoxigenic Escherichia coli (ETEC) isolates (n = 170) obtained from preweaning piglet colibacillosis from the two different pig production systems (commercial piggeries and small holder farmers) in Vietnam. Overall, tetracycline, streptomycin, amoxicillin, trimethoprim/sulfamethoxazole, and chloramphenicol showed markedly higher rates of resistance. Both apramycin and ceftiofur are active against all ETEC isolates. These antimicrobials could be recommended as the drugs of choice for the treatment of E. coli infections in young pigs in North Vietnam. Resistance to third-generation cephalosporin (ceftiofur, ceftazidime, and cefoxitin) was not observed in Vietnamese ETEC isolates. Multiple resistances to greater than three antimicrobials were widely distributed (approximately 79.4%).

Animal Husbandry↗

Comparison of essential oils from three plants for enhancement of antimicrobial activity of nitrofurantoin against enterobacteria.

BACKGROUND: Piperitone from plant essential oils enhancesbactericidal activities of nitrofurantoin and furazolidone against bacteria from the family Enterobacteriaceae. In this study, the essential oils of spearmint (Mentha spicata L.), dill (Anethum graveolens L.) and peppermint (Mentha piperita L.)were screened for augmentation of nitrofurantoin activity and the most active components were determined. METHOD: The effects of essential oils and their components on the bactericidal activity of nitrofurantoin against Enterobacter cloacae were studied using disk-diffusion and agar-dilution methods. The composition of essential oils was studied using gas chromatography-mass spectrometry. RESULTS: M. spicata and A. graveolens oils exhibited the highest effects. Gas chromatography-mass spectrometry analysis showed that the oils of these two plants contained 40.12 and 20.32% carvone, respectively. Pure carvone and piperitone equally increased the bactericidal activity of nitrofurantoin. Other ingredients of essential oils, including camphor, limonene and menthone, were less effective.

Anethum graveolens↗